Ornamental kale, as a burgeoning landscaping plant, is gaining popularity for its rich color patterns in leaf and cold tolerance. Leaf variegation endows ornamental kale with unique ornamental characters, and the mutants are ideal materials for exploring the formation mechanisms of variegated phenotype. Herein, we identified a novel variegated leaf kale mutant ‘JC007-2B’ with green margins and white centers. Morphological observations and physiological determinations of the green leaf stage (S1), albino stage (S2) and variegated leaf stage (S3) demonstrated that the chloroplast structure and photosynthetic pigment content in the white sectors (S3_C) of variegated leaves were abnormal. Genetic analysis revealed that a single dominant nuclear gene (BoVl) controlled the variegated leaf trait of ‘JC007-2B’, and three candidate genes for BoVl were fine-mapped to a 6.74 Kb interval on chromosome C03. Multiple sequence alignment among the green-leaf mapping parent ‘BS’, recombinant individuals, mutant parent ‘JC007-2B’ and its same originated DH line population established that the mutation sites in Bo3g002080 exhibited a complete consensus. Bo3g002080, homologous to Arabidopsis MED4, was identified as the candidate gene for BoVl. Expression analysis showed that Bo3g002080 displayed a 2158.85-fold higher expression at albino stage than that in green leaf stage. Transcriptome analysis showed that related pathways of photosynthesis and chloroplast development were significantly enriched in the white sectors, and relevant DEGs involved in these pathways were almost down-regulated. Overall, our study provides a new gene resource for cultivar breeding in ornamental kale and contributes to uncovering the molecular genetic mechanism underlying the variegated leaf formation.

/pdf/fine-mapping-of-bovl-conferring-the-variegated-leaf-in-194myi67.pdf

Fine Mapping of BoVl Conferring the Variegated Leaf in Ornamental Kale (Brassica oleracea var. acephala)

Ilex × attenuata ‘Sunny Foster’ represents a yellow leaf mutant originating from I. × attenuata ‘Foster#2’, a popular ornamental woody cultivar. However, the molecular mechanisms underlying this leaf color mutation remain unclear. Using a comprehensive approach encompassing cytological, physiological, and transcriptomic methodologies, notable distinctions were discerned between the mutant specimen and its wild type. The mutant phenotype displayed aberrant chloroplast morphology, diminished chlorophyll content, heightened carotenoid/chlorophyll ratios, and a decelerated rate of plant development. Transcriptome analysis identified differentially expressed genes (DEGs) related to chlorophyll metabolism, carotenoid biosynthesis and photosynthesis. The up-regulation of CHLD and CHLI subunits leads to decreased magnesium chelatase activity, while the up-regulation of COX10 increases heme biosynthesis—both impair chlorophyll synthesis. Conversely, the down-regulation of HEMD hindered chlorophyll synthesis, and the up-regulation of SGR enhanced chlorophyll degradation, resulting in reduced chlorophyll content. Additionally, genes linked to carotenoid biosynthesis, flavonoid metabolism, and photosynthesis were significantly down-regulated. We also identified 311 putative differentially expressed transcription factors, including bHLHs and GLKs. These findings shed light on the molecular mechanisms underlying leaf color mutation in I. × attenuata ‘Sunny Foster’ and provide a substantial gene reservoir for enhancing leaf color through breeding techniques.

Molecular Mechanisms of Chlorophyll Deficiency in Ilex × attenuata ‘Sunny Foster’ Mutant

he formation of chimera is a well-known phenomenon in variegated ornamental and wild plants, but the biochemical peculiarities of this phenomenon are not well studied. The purpose of this work was to conduct a comparative study of the photosynthetic apparatus functioning, the content of macro-, microelements, flavonoids and brassinosteroids in the variegated and green leaves of the periclinal chimera Cornus controversa plant grown in the collection of M. M. Gryshko National Botanical Garden of the NAS of Ukraine. Sampling was made during the period from June to October 2022. The content of photosynthetic pigments and brassinosteroids was measured spectrophotometrically, autofluorescence of leaf tissues was studied using an inverted microscope with a multi-channel fluorescence imaging system, Normalized difference vegetation index (NDVI) – using GreenSeeker devices and fluorimeter, the level of macro- and microelements – using plasma emission spectrometer. Phenolic secondary metabolites were separated by HPTLC. To assess the protective capacity of flavonoids, a bioassay on the germination of radish seeds Raphanus in the presence of AlCl3 was used. It was shown that green leaves of C. controversa chimera, compared to variegated ones, had a twice as high NDVI index, maintained photosynthetic function longer in autumn, and had higher content of macronutrients (Mg, Ca and S) but lower concentration of toxic metals (Al, Cu, Fe, Zn, Ti). Instead, variegated leaves accumulated more brassinosteroids and phenolic metabolites, which were found not only to reduce the toxic effect of aluminum but also to have a growth-stimulating effect on the roots of radish seedlings. Such specialization of green and variegated leaves may contribute to better plant chimeras adaptation to environmental pollution. Keywords: brassinosteroids, Cornus controversa, macroelements, microelements, NDVI index, periclinal chimera, phenolic metabolites, photosynthetic pigments, tissue autofluorescence 

Photosystems functioning, the content of chemical elements, flavonoids and brassinosteroids in leaves of chimera Cornus controversa

Abstract Background Primulina pungentisepala is suitable for use as a potted plant because of its beautiful leaf variegation, which is significantly different in its selfed offspring. However, the mechanism of P. pungentisepala leaf variegation is unclear. In this study, two types of offspring showing the greatest differences were compared in terms of leaf structure, chlorophyll contents, chlorophyll fluorescence parameters and transcriptomes to provide a reference for studying the molecular mechanism of structural leaf variegation. Results Air spaces were found between water storage tissue, and the palisade tissue cells were spherical in the white type. The content of chlorophyll a and total chlorophyll (chlorophyll a + b) was significantly lower in the white type, but there were no significant differences in the content of chlorophyll b, chlorophyll a/b or chlorophyll fluorescence parameters between the white and green types. We performed transcriptomic sequencing to identify differentially expressed genes (DEGs) involved in cell division and differentiation, chlorophyll metabolism and photosynthesis. Among these genes, the expression of the cell division- and differentiation-related leucine-rich repeat receptor-like kinases (LRR-RLKs), xyloglucan endotransglycosylase/hydrolase ( XET/H ), pectinesterase ( PE ), expansin ( EXP ), cellulose synthase-like ( CSL ), VARIEGATED 3 ( VAR3 ), and ZAT10 genes were downregulated in the white type, which might have promoted the development air spaces and variant palisade cells. Chlorophyll biosynthesis-related hydroxymethylbilane synthase ( HEMC ) and the H subunit of magnesium chelatase ( CHLH ) were downregulated, while chlorophyll degradation-related chlorophyllase-2 ( CHL2 ) was upregulated in the white type, which might have led to lower chlorophyll accumulation. Conclusion Leaf variegation in P. pungentisepala was caused by a combination of mechanisms involving structural variegation and low chlorophyll levels. Our research provides significant insights into the molecular mechanisms of structural leaf variegation. 

/pdf/cytological-physiological-and-transcriptomic-analysis-of-3p4wzezy.pdf

Cytological, physiological and transcriptomic analysis of variegated Leaves in Primulina pungentisepala offspring

/pdf/cytological-physiological-and-transcriptomic-analysis-of-qqkk9seq.pdf

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Cytological, physiological and transcriptomic analysis of variegated Leaves in Primulina pungentisepala offspring

Cytological, physiological and transcriptomic analysis of variegated Leaves in Primulina pungentisepala offspring